The present invention relates to antibody substances and more particularly to antibody substances displaying unique, multi-specific immunoreactivities with respect to glycoprotein D of Herpes Simplex Virus types 1 and 2 (HSV gD-1 and gD-2) and structurally related compounds.
In the recent past, substantial research efforts have been directed to the isolation and characterization of the Herpes Simplex Virus glycoproteins which constitute structural components of the virion envelope and are likely to play an important role in initiation of viral infection. Among the more significant advances based upon this research is the discovery by the present applicants of novel preparations of HSV glycoprotein D which, when employed as the active immunogen of vaccine compositions, provoke significant protection against HSV infection and applicants further discovery of immunologically significant polypeptides which duplicate or substantially duplicate continuous sequences of amino acids extant in HSV gD. Incorporated by reference herein for the purpose of providing background information with respect to the art are the disclosures of applicants' co-pending U.S. Patent Application Ser. No. 463,141, filed Feb. 4, 1983, and entitled "Methods and Materials for Herpes Simplex Virus Vaccination". (See, also, International Patent Application W083/02897, published Sept. 1, 1983.)
Research in characterization of HSV gD-1 and gD-2 has been substantially assisted by the results obtained through application of recombinant DNA techniques to the cloning and expression in microbial hosts (e.g., bacteria, yeast and mammalian cells in culture) of DNA sequences coding for part or all of the polypeptide sequences of gD-1 and gD-2. DNA sequencing of genes coding for the proteins has resulted in prediction of their primary structural conformation (amino acid sequence). See, e.g., Watson, et al., Science, 218, pp. 381-384 (1982) providing the predicted sequence for gD-1 including both a "leader" region and a region designated as that of the "mature" protein. Similar recombinant methods have allowed the prediction of the gD-2 sequence and the comparison thereof with gD-1. Also specifically incorporated herein by reference, therefore, are the disclosures of Watson, Gene, 26, pp. 307-312 (1983) which generally provide information as set forth below in Table I concerning the comparative primary structural conformation (amino acid sequences) predicted for "mature" HSV gD-1 and gD-2 based on DNA sequencing.
In the Table and throughout, the following single and triple letter "codes" for amino acid residues will be employed: A=Ala=Alanine; C=Cys=Cysteine; D=Asp=Aspartic Acid; E=Glu=Glutamic Acid; F=Phe=Phenylalanine; G=Gly=Glycine; H=His=Histidine; I=Ile=Isoleucine; K=Lys=Lysine; L=leu=Leucine; M=Met=Methionine; N=Asn=Asparagine; P=Pro=Proline; Q=Gln=Glutamine; R=Arg=Arginine; S=Ser=Serine; T=Thr=Threonine; V=Val=Valine; W=Trp=Tryptophan; and Y=Tyr=Tyrosine.
TABLE I __________________________________________________________________________ 1 gD-1 KYALADA SLKMADPNRFRGKD LPVLDQLTDPPGVRRVYHI 40 gD-2 P---------------- ----------------N--------------- ---------------K ----------- gD-1 QAGLPDPFQPPSLPIT VYYAVLERACRSV LLNAPSEAPQI 80 gD-2 PS--E---------------I--------------- --------------- ----------H---------- ------- gD-1 VRGASEDVRKQP YNLTIAWFRMGGNCA IPITVMEYTECSY 120 gD-2 DEA----HT ----------------Y --------D---------------- ---------------P-- gD-1 NKSLGACPIRTQPRWN YYDSFSAVSEDNLGFLMHAPAFET 160 gD-2 V ---------- ----------S--------------- --------------- --------------- ----------- gD-1 AGTYLRLVKINDWTEIT QFILEHRAKGSCK YALPLRIPPS 200 gD-2 RA----- --------------- -----A gD-1 ACLSPQYQQGVTVDSIGM LPRFIPENQRTVAVYSLKIAG 240 gD-2 TSK ---------------------------------------------------------L------------ --- gD-1 WHGPKAPYTSTLLPPEL SETPNATQPELA PEDPEDSALLE 280 gD-2 P------------------------D-T---------- --------V --------------- ---------- gD-1 DPVGTVAPQIPPNWHIPSIQDAATP YHPPATPNNMGLIAG 320 gD-2 A-------SS---------------------------V------H--A----A--S-P------I---- 319 gD-1 AVGGSLLAALVICGIVY WMHRRTRKAPKRI RLPHIREDDQ 360 gD-2 LA----T------------- G----AF---VR-----AQM---------L------------ 359---A gD-1 PSSHQPLFY 369 gD-2 P--------------- 368 __________________________________________________________________________
Briefly summarized, mature forms of gD-1 and gD-2 are predicted to consist of 369 and 368 amino acid residues, respectively, with gD-2 "lacking" a residue corresponding to residue 304 of gD-1, and with approximately 85% homology existing between the two sequences. See, also, Lasky, et al., DNA, 3, pp. 23-29 (1984) and Rawls, et al., J.Virol., 51, pp. 263-265 (1984).
Companion research in the characterization of HSV gD-1 and gD-2 has been directed to the localization of antigenic determinants of these materials [see, Eisenberg, et al., J.Virol., 41, pp. 1099-1104 (1982), Cohen, et al., J.Virol., 49, pp. 102-108 (1984), Eisenberg, et al., J.Virol., 49, pp. 265-268 (1984) and references cited therein] through the use of typespecific and type-common monoclonal antibody substances. The essentially dual goals of this work have been the ascertaining of one or more determinants which stimulate production of neutralizing antibodies in HSV hosts and the identification of antibody substances correspondingly useful in detection, quantification, and affinity purification of such immunologically significant substances.
Despite the above-noted advances in the art, there continues to exist a need for antibody substances useful in the detection, quantification and isolation by affinity purification from natural and recombinant sources of gD-1, gD-2 and structurally related compounds such as gD-1 and gD-2 fragments and/or analogs.